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Per‐glycosylation of the Surface‐Accessible Lysines: One‐Pot Aqueous Route to Stabilized Proteins with Native Activity
Author(s) -
Walther Raoul,
Monge Pere,
Pedersen Andreas Bøtker,
Benderoth Anja,
Pedersen Jannik Nedergaard,
Farzadfard Azad,
Mandrup Ole A.,
Howard Kenneth A.,
Otzen Daniel E.,
Zelikin Alexander N.
Publication year - 2021
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.202100228
Subject(s) - glycosylation , biochemistry , chemistry , recombinant dna , computational biology , biology , gene
Chemical glycosylation of proteins is a powerful tool applied widely in biomedicine and biotechnology. However, it is a challenging undertaking and typically relies on recombinant proteins and site‐specific conjugations. The scope and utility of this nature‐inspired methodology would be broadened tremendously by the advent of facile, scalable techniques in glycosylation, which are currently missing. In this work, we investigated a one‐pot aqueous protocol to achieve indiscriminate, surface‐wide glycosylation of the surface accessible amines (lysines and/or N‐terminus). We reveal that this approach afforded minimal if any change in the protein activity and recognition events in biochemical and cell culture assays, but at the same time provided a significant benefit of stabilizing proteins against aggregation and fibrillation ‐ as demonstrated on serum proteins (albumins and immunoglobulin G, IgG), an enzyme (uricase), and proteins involved in neurodegenerative disease (α‐synuclein) and diabetes (insulin). Most importantly, this highly advantageous result was achieved via a one‐pot aqueous protocol performed on native proteins, bypassing the use of complex chemical methodologies and recombinant proteins.